1.Inhibition of MGMT gene by hairpin siRNA expression vector in vitro
Fang WANG ; Ziyuan ZHOU ; Shengxue LIU ; Jia CAO
Journal of Third Military Medical University 2003;0(11):-
Objective To construct the small interfering RNA (siRNA) eukaryotic expression vector specific to human MGMT gene(pRNATin-H1.2/Neo MGMT siRNA) to observe its silencing effect on MGMT gene in vitro.Methods The pRNATin-H1.2/Neo MGMT siRNA expression vector was constructed by gene recombination,then transfected into the cultured HelaS3 cells.Inhibitory effect of siRNAs was detected by semi-quantitative RT-PCR.Results The pRNATin-H1.2/Neo MGMT siRNA expression vector was successfully constructed.Cells transfected with pRNATin-H1.2/Neo MGMT siRNA could obviously inhibit the expression level of MGMT gene.Conclusion The pRNATin-H1.2/Neo MGMT siRNA expression vector could inhibit the MGMT gene expression.
2.Alteration levels of IL-17 and IL-19 inflammatory factors among deadaptation personnel from the plateau
Xingming QU ; Binfeng HE ; Hongping CHEN ; Shengxue SHI ; Guisheng QIAN ; Guansong WANG
Military Medical Sciences 2014;(10):768-770,779
Objective To analyze the change in the concentration of IL-17 and IL-10 inflammatory factors among the deadaptation personnel who returned from the plateau.Methods A total 21 healthy males were investigated who averaged 25 years in age, lived permanently in the plains (200 m), and once stayed to the plateau (Lasha) for 6 months.Their venous blood was collected at three time points:the day before ascending to the plateau(control), the second day after return to the plains(d2) and the 30th day(d30), respectively.Their serum was seperated from the whole blood and the level of IL-17A and IL-10 was detected by ELISA method.Results The concentration of IL-17A and the IL-17A/IL-10 ratio were significant increased at d2 and d30, respectively, compared with control (P<0.05).Compared with d2, IL-17A and the IL-17A/IL-10 ratio were decreased obviously at d30(P <0.05).The level of IL-10 at d2 and d30 was significantly reduced compared with control ( P <0.05), but increased at d30 compared with d2.Correlative analysis showed that there was a negative correlation in the levels of IL-10 and IL-17A between control, d2 and d30, respectively (r1=0.948, P<0.05;r2=0.969, P<0.05;r3=0.972, P<0.05).A significant negative correlation was observed in the alteration levels of IL-10 and IL-17A between the three groups(r4=-0.793, P<0.05; r5=-0.756, P<0.05). Conclusion The concentration of inflammatory factors among the plateau deadaptation patients is imbalanced, but it is gradually reduced with time.The mechanism is still not clear.
3.Expression of synapse-related proteins in hippocampus of rats with epilepsy accompanied by depression
Hongmei QIU ; Wang GUO ; Xueping YANG ; Shengxue WANG ; Yuhao CHU ; Yun LI
Chinese Journal of Behavioral Medicine and Brain Science 2024;33(1):9-14
Objective:To investigate the expression of hippocampal synapse-related proteins including synaptophysin (SYN), postsynaptic density protein 95 (PSD95) and growth-associated protein 43 (GAP43) in rats with epilepsy accompanied by depression.Methods:The 3-month-old female clean grade SD rats were selected for the experiment.Lithium chloride pilocarpine was used to establish an epileptic rat model. Rats with successful epilepsy models were divided into epileptic depressive group (EWD group)and epileptic group with 10 in each group based on whether they were accompanied by depression. Furthermore, ten rats with matched body mass were taken as the depressive group and 10 were taken as control group. As for the depressive group rats, chronic unpredictable mild stress stimulation combined with orphanage was adopted to establish a model of depression.The depressive behaviors of rats were evaluated by body mass, sucrose preference test and open field test. Immunohistochemical staining and Western blot were used to detect the expression of SYN, PSD95 and GAP43 proteins in rat hippocampal tissue. SPSS 17.0 software was used for data statistical analysis, repeated measurement ANOVA was used for behavioral results, one-way ANOVA was used for inter group comparison of protein expression data, and LSD test was used for further pairwise comparison.Results:As for the body mass, there was significant interaction effect between the time and group among the 4 groups ( F=7.33, P<0.01). On the 8th day and the 29th day, the body weight of rats in the EWD group and the depressive group were lower than those in the epilepsy group (all P<0.05). The body weight of rats in the EWD group on the 29th day was lower than that on the first day ( P<0.05). As for the sucrose preference rates, there was significant interaction effect between the time and group among the 4 groups( F=2.67, P<0.05). The sucrose preference rate of EWD group on the15th and 29th day were lower than that on the first day (both P<0.05). The results of the open field test showed that the interaction effects of the number of vertical standing times( F=2.74) and the number of horizontal movement lattices ( F=1.76) both were not significant (both P>0.05), but both the time effect and group effect were significant (vertical standing times: Ftime=4.35, P<0.05, Fgroup=25.64, P<0.01; horizontal movement lattices: Ftime=12.75, P<0.01, Fgroup=21.37, P<0.01). The immunohistochemical results showed that there was a statistically significant difference in the number of positive cells expressing synaptic proteins SYN, PSD95 and GAP43 among the four groups of rats ( F=93.85, 58.66, 98.84, all P<0.05). The numbers of positive cells of SYN (11.73±4.30), PSD95 (24.47±7.58) and GAP43 (9.40±3.50) in the epilepsy group were lower than those in the control group ((51.00±15.39), (55.60±13.17) and (29.53±4.05)) (all P<0.05). The numbers of positive cells of SYN (5.80±3.53), PSD95 (12.87±4.03) and GAP43 (5.33±3.50) in the EWD group were lower than those in the depressive group ((11.33±3.22), (48.13±12.69) and (15.47±5.21) )(all P<0.05). Western blot results showed that there were statistically significant differences in the expression of synaptic proteins SYN, PSD95 and GAP43 among the four groups of rats ( F=13.19, 9.38, 16.80, all P<0.05). The expression levels of SYN, PSD95 and GAP43 in the epilepsy group were lower than those in the control group (all P<0.05). The expression levels of SYN, PSD95 and GAP43 in the EWD group were lower than those in the epilepsy group and the depressive group (all P<0.05). Conclusion:The low expression of SYN, PSD95 and GAP43 proteins in the hippocampus of rats with epilepsy accompanied by depression may be related to their pathogenesis.
4.Development and validation of a nomogram model for preoperative prediction of hepatocellular carcinoma with microvascular invasion
Kangkang WAN ; Shubo PAN ; Liangping NI ; Qiru XIONG ; Shengxue XIE ; Longsheng WANG ; Tao LIU ; Haonan SUN ; Ju MA ; Huimin WANG ; Zongfan YU
Chinese Journal of Hepatobiliary Surgery 2023;29(8):561-566
Objective:To develop and validate a nomogram model for predicting microvascular invasion (MVI) in hepatocellular carcinoma (HCC) based on preoperative enhanced computed tomography imaging features and clinical data.Methods:The clinical data of 210 patients with HCC undergoing surgery in the Second Affiliated Hospital of Anhui Medical University from May 2018 to May 2022 were retrospectively analyzed, including 172 males and 38 females, aged (59±10) years old. Patients were randomly divided into the training group ( n=147) and validation group ( n=63) by systematic sampling at a ratio of 7∶3. Preoperative enhanced computed tomography imaging features and clinical data of the patients were collected. Logistic regression was conducted to analyze the risk factors for HCC with MVI, and a nomogram model containing the risk factors was established and validated. The diagnostic efficacy of predicting MVI status in patients with HCC was assessed by receiver operating characteristic (ROC) curve, calibration curves, decision curve analysis (DCA), and clinical impact curve (CIC) of the subjects in the training and validation groups. Results:The results of multifactorial analysis showed that alpha fetoprotein ≥400 μg/ml, intra-tumor necrosis, tumor length diameter ≥3 cm, unclear tumor border, and subfoci around the tumor were independent risk factors predicting MVI in HCC. A nomogram model was established based on the above factors, in which the area under the curve (AUC) of ROC were 0.866 (95% CI: 0.807-0.924) and 0.834 (95% CI: 0.729-0.939) in the training and validation groups, respectively. The DCA results showed that the predictive model thresholds when the net return is >0 ranging from 7% to 93% and 12% to 87% in the training and validation groups, respectively. The CIC results showed that the group of patients with predictive MVI by the nomogram model are highly matched with the group of patients with confirmed MVI. Conclusion:The nomogram model based on the imaging features and clinical data could predict the MVI in HCC patients prior to surgery.
5.Clinical evaluation of two different peritoneal dialysis catheter removal methods in elderly patients on peritoneal dialysis
Shengxue ZHANG ; Xiaole SU ; Yan YAN ; Yan ZHAO ; Xinyan LIU ; Xiaoxia LI ; Yan ZHANG ; Lihua WANG
Chinese Journal of Geriatrics 2022;41(2):191-195
Objective:To assess two methods for peritoneal dialysis catheter removal in elderly patients on peritoneal dialysis.Methods:A retrospective cohort study was conducted and clinical data were collected for 107 elderly patients on peritoneal dialysis with catheters removed at the peritoneal dialysis center of the Second Hospital of Shanxi Medical University from August 2010 to May 2020.The 107 patients were divided into the open surgical extubation group and the 'pull' technique group.Sex, age, disease leading to dialysis, dialysis age, reason for extubation and preoperative test results were compared between the two groups.Operative time, length of postoperative hospitalization, perioperative pain and postoperative complications of the two groups were examined.Results:Higher levels or values were observed in operative time[(71.2±13.4)min vs.(19.3±5.6)min, t=16.933, P<0.01], length of postoperative hospitalization[(9.5±1.8)d vs.(2.2±0.5)d, t=10.988, P<0.01]and intraoperative pain score(4.4±1.6 vs.1.4±1.1, t=6.909, P<0.01)and pain score 24h after surgery(3.7±1.4 vs.0.5±0.3, t=9.995, P<0.01)in the open surgical extubation group than in the 'pull' technique group, but there was no statistical difference in the incidence of postoperative complications between the two groups(6.8% vs.5.0%, χ2=0.037, P>0.05). Conclusions:either the open surgical removal method or 'pull' technique.can safely and effectively remove the peritoneal dialysis catheter in elderly patients on peritoneal dialysis of the two methods, the 'pull' technique has advantages including less trauma, shorter and simpler surgery, helping achieve improved quality of life for elderly patients on peritoneal dialysis.
6.Correlation between pathological types of parathyroid and clinical manifestations in patients with renal secondary hyperparathyroidism
Yali ZHAN ; Xiaohe WANG ; Deguang WANG ; Sen ZHANG ; Yang WAN ; Jifeng WU ; Shengxue XIE ; Li HAO
Chinese Journal of Nephrology 2018;34(3):179-184
Objective To analyze the correlation between the pathological types of parathyroid and clinical manifestations in patients with renal secondary hyperparathyroidism (SHPT),so as to improve the efficacy and safety of treatment.Methods The pathological and clinical data of 130 patients with renal SHPT and maintenance hemodialysis (MHD) who had undergone total parathyroidectomy with autotransplantation (TPTX + AT) were collected.A total of 545 parathyroid glands were obtained and 998 slices were made and read.According to the pathological types of parathyroid hyperplasia,the patients were divided into diffuse hyperplasia (DH) group,diffuse between hyperplasia and nodular hyperplasia (DH/NH) group as well as nodular hyperplasia (NH) group.The clinical and biochemical characteristics of different groups before and after operation (1-,3-,6-,9-,12-month) were compared and analyzed by statistical tests.Results (1) The preoperative status:the dialysis age,serum calcium as well as incidence of bone pain,skin itching and shorten height in the NH group were significantly higher than those in the DH group (all P < 0.05),and the serum phosphorus and iPTH in the NH group were significantly higher than those in DH and DH/NH group (all P < 0.05).(2) The postoperative status:the serum calcium of the NH group at 1-month was lower than that of the DH group,and the incidence of hypocalcemia of the NH group at 1-month was higher than that of the DH group (P < 0.05);the serum phosphorus at 3-,6-,9-month and iPTH at 1-,3-month of the NH group were significantly lower than that of the DH group (all P < 0.05),and the serum phosphorus at 3-month and iPTH at 1-month of the NH group were lower than that of the DH/NH group (all P < 0.05).Among the 3 groups the serum phosphorus change from 1 to 12 months had difference (F=3.241,P=0.042),while the differences of serum calcium and iPTH changes were statistically insignificant.Conclusions The clinical manifestations,serum calcium,phosphorus and iPTH in patients with renal SHPT before and after TPTX+AT are closely related to the pathological types of parathyroid hyperplasia.Compared with the DH patients,before the operation the NH patients have longer dialysis age,more serious the clinical symptoms such as bone disease,higher calcium,phosphorus and iPTH,while greater reduction of the serum calcium,phosphorus and iPTH in the short term after operation.
7.Development of a
Yiying YANG ; Qingqing SUN ; Yang LIU ; Hanzhi YIN ; Wenping YANG ; Yang WANG ; Ying LIU ; Yuxian LI ; Shen PANG ; Wenxi LIU ; Qian ZHANG ; Fang YUAN ; Shiwen QIU ; Jiong LI ; Xuefeng WANG ; Keqiang FAN ; Weishan WANG ; Zilong LI ; Shouliang YIN
Journal of Zhejiang University. Science. B 2021;22(5):383-396